Sreekumar Parameswaran G, Zhou Jiehao, Sohn Joonhong, Spee Christine, Ryan Stephen J, Maurer Barry J, Kannan Ram, Hinton David R
The Arnold and Mabel Beckman Macular Research Center, Doheny Eye Institute, Childrens Hospital of Los Angeles, Keck School of Medicine, University of Southern California, Los Angeles, CA 90033, USA.
Invest Ophthalmol Vis Sci. 2008 Mar;49(3):1210-20. doi: 10.1167/iovs.07-0667.
To evaluate the effect of N-4-hydroxyphenyl retinamide (4-HPR) on experimental laser-induced choroidal neovascularization (CNV) and on the expression and secretion of relevant growth factors by cultured human retinal pigment epithelial (RPE) cells.
CNV was induced by laser photocoagulation in C57BL/6 mice. 4-HPR (0.2 or 1 mg) or vehicle, was injected intraperitoneally twice daily for 14 days. Plasma and tissue levels of 4-HPR were measured by HPLC. CNV was evaluated by fluorescein angiography, histology, and quantitative confocal analysis of isolectin B4 histochemistry on days 7 and 14. Induction of apoptosis and expression and secretion of growth factors was studied in 4-HPR-treated RPE cultures.
Mice treated with 4-HPR exhibited time- and dose-dependent increases in plasma and tissue 4-HPR levels. CNV lesions showed increased volume with increased vascular leakage and contained fewer lesion-associated RPE in treated versus untreated mice. Treatment of nonpolarized RPE cultures with 4-HPR in the presence of serum resulted in RPE apoptosis; however, apoptosis was minimal in similarly treated highly polarized RPE. Treatment of RPE cells with 4-HPR resulted in the upregulation of VEGF-A and -C (P < 0.05) and Ang-1 (P < 0.01) mRNA and increased secretion of VEGF-A and -C (P < 0.05), whereas pigment epithelium-derived growth factor (PEDF) and thrombospondin (TSP)-1 mRNA expression and secretion were downregulated (P < 0.05).
4-HPR increases lesion size and leakage in laser-induced CNV and is associated with the upregulation of key proangiogenic factors and the downregulation of antiangiogenic factors. Consistent with the preferential loss of RPE in CNV lesions in vivo, 4-HPR induces apoptosis of nonpolarized RPE in the presence of serum.
评估N - 4 - 羟基苯基视黄酸（4 - HPR）对实验性激光诱导脉络膜新生血管（CNV）的影响，以及对培养的人视网膜色素上皮（RPE）细胞相关生长因子表达和分泌的影响。
通过激光光凝在C57BL / 6小鼠中诱导CNV。4 - HPR（0.2或1毫克）或赋形剂，每天腹腔注射两次，共14天。通过高效液相色谱法测量血浆和组织中4 - HPR的水平。在第7天和第14天，通过荧光素血管造影、组织学以及异凝集素B4组织化学的定量共聚焦分析评估CNV。在4 - HPR处理的RPE培养物中研究细胞凋亡的诱导以及生长因子的表达和分泌。
用4 - HPR处理的小鼠血浆和组织中4 - HPR水平呈现时间和剂量依赖性增加。与未处理的小鼠相比，处理组的CNV病变体积增加，血管渗漏增加，且病变相关的RPE细胞减少。在血清存在的情况下，用4 - HPR处理非极化的RPE培养物会导致RPE细胞凋亡；然而，在类似处理的高度极化的RPE中，细胞凋亡极少。用4 - HPR处理RPE细胞导致VEGF - A和 - C（P < 0.05）以及Ang - 1（P < 0.01）mRNA上调，VEGF - A和 - C的分泌增加（P < 0.05），而色素上皮衍生生长因子（PEDF）和血小板反应蛋白（TSP）-1 mRNA表达及分泌下调（P < 0.05）。
4 - HPR增加激光诱导的CNV病变大小和渗漏，并与关键促血管生成因子上调和抗血管生成因子下调相关。与体内CNV病变中RPE的优先丢失一致，4 - HPR在血清存在的情况下诱导非极化RPE细胞凋亡。
